Chulalongkorn University Dental Journal

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Objective To 0.07develop a PCR-based method for the detection and identification of Candida albicans and C. glabrata in oral samples, with a comparison of sample preparations by DNA extraction and boiling. Materials and methods Oral rinse samples were collected from 10 healthy volunteers. A portion of the samples was used for cultivation and species identification by growth on Chromogenic Candida agar, chlamydospore formation and biochemical assays. The second portion was subjected to DNA extraction, while the third portion was washed and boiled for 10 minutes. The samples were than tested by PCR using fungal specific primers (ITS) and species specific primers for C. albicans (CALB) and C. glabrata (CGLA). PCR products were detected by standard agarose gel electrophoresis. Results Using culture-based assays, Candida was detected in 7 samples, all of which contain C. albicans while 2 also contain C. albicans while 2 also contain C. glabrata. In DNA extracted samples, the PCR assay using ITS primers detected fungi in 6 samples (85.71% sensitivity, 100% specificity); CALB primers detected C. albicans in 5 samples (71.43% sensitivity, 100% specificity), and CGLA primers detected C. glabrata in 1 of the 2 samples. This PCR assay was unable to detect the presence of Candida in boiled samples. Conclusion This PCR assay was able to detect and identify C. albicans and C. glabrata with high specificity in DNA extracted oral samples, but not in boiled samples. Therefore, the simple and economical DNA extracted method used in this study can be applied for the rapid detection and identification of Candida in oral samples by PCR.



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